Many origins-of-life scenarios depict a situation in which there are common and potentially scarce resources needed by molecules that compete for survival and reproduction. The dynamics of RNA assembly in a complex mixture of sequences is a frequencydependent process and mimics such scenarios. By synthesizing Azoarcus ribozyme genotypes that differ in their single-nucleotide interactions with other genotypes, we can create molecules that interact among each other to reproduce. Pairwise interplays between RNAs involve both cooperation and selfishness, quantifiable in a 2 × 2 payoff matrix. We show that a simple model of differential equations based on chemical kinetics accurately predicts the outcomes of these molecular competitions using simple rate inputs into these matrices. In some cases, we find that mixtures of different RNAs reproduce much better than each RNA type alone, reflecting a molecular form of reciprocal cooperation. We also demonstrate that three RNA genotypes can stably coexist in a rock-paper-scissors analog. Our experiments suggest a new type of evolutionary game dynamics, called prelife game dynamics or chemical game dynamics. These operate without templatedirected replication, illustrating how small networks of RNAs could have developed and evolved in an RNA world.RNA | prebiotic chemistry | origins of life | game theory | ribozyme A plausible description of a sequence of events that could have led to the origins of life on the Earth from a purely chemical milieu has long been desirable, yet remains elusive. The RNA world hypothesis has helped sharpen our focus on what could have taken place 4 Gya, in that RNA serves as a powerful model for a self-sustaining chemical system capable of evolutionary change (1-6). Although this hypothesis has engendered much debate, both in its general applicability and in the details of its implementation (7-9), there are some clear emerging trends. Among the recent advances in prebiotic RNA studies is the concept of an evolving "network" of RNAs being required to kick-start life, rather than a single selfish entity. This idea dates back to the formative studies of Eigen and Schuster in the 1970s (10, 11). However, it can be sharply seen in the 20+-y effort aimed at developing a generalized RNA replicase ribozyme in the laboratory: new successes have taken advantage of a fragmentation of the best such artificial ribozyme and invoke a network of reactions to provide for its assembly (12). Our own laboratories have focused on a variety of "prelife" (13, 14) and cooperative network (15, 16) approaches to understand how evolving RNA systems could have arisen from abiotic sources of nucleotides and short oligomers. Many others have also stressed the need for distributed functionality at the onset of life, both chemically (17) and in space and time (18,19).To advance a network approach to the "single biomolecule problem" in the RNA world, what is needed now is an understanding of how prebiotic networks could have evolved. Auspiciously, the mechanisms of network evol...